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Brown Adipogenic Reprogramming Induced by a Small Molecule

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Presentation on theme: "Brown Adipogenic Reprogramming Induced by a Small Molecule"— Presentation transcript:

1 Brown Adipogenic Reprogramming Induced by a Small Molecule
Baoming Nie, Tao Nie, Xiaoyan Hui, Ping Gu, Liufeng Mao, Kuai Li, Ran Yuan, Jiashun Zheng, Haixia Wang, Ke Li, Shibing Tang, Yu Zhang, Tao Xu, Aimin Xu, Donghai Wu, Sheng Ding  Cell Reports  Volume 18, Issue 3, Pages (January 2017) DOI: /j.celrep Copyright © 2017 The Authors Terms and Conditions

2 Cell Reports 2017 18, 624-635DOI: (10.1016/j.celrep.2016.12.062)
Copyright © 2017 The Authors Terms and Conditions

3 Figure 1 High-Throughput Screening Identified Bex, which Induces Brown Adipogenic Reprogramming of Committed Myoblasts (A) Schematic of the high-throughput chemical screening for brown adipogenic reprogramming of committed myoblasts. (B) Bex-induced BAT-like adipocytes from C2C12 myoblasts. Lipid droplets were stained by oil red O. (C) qPCR analysis of BAT genes in BAT-like adipocytes from C2C12 myoblasts. Data were normalized to Ctrl and represented mean ± SEM. ∗∗p < 0.01, ∗∗∗p < (n = 3). (D) UCP1 immunostaining in Bex-induced BAT-like adipocytes from C2C12 myoblasts. Cell Reports  , DOI: ( /j.celrep ) Copyright © 2017 The Authors Terms and Conditions

4 Figure 2 RXRα/γ Agonism Mediates Bex-Induced Brown Adipogenic Reprogramming (A) The RXR antagonist HX531 inhibits Bex-induced brown adipogenic reprogramming of myoblasts. (B) Knockdown of Rxr subtypes by shRNAs targeting Rxrα, Rxrβ, or Rxrγ abolished Bex-induced brown adipogenic reprogramming of myoblasts. (C) Overexpression of Rxrα and Rxrγ, but not Rxrβ, dramatically potentiated Bex-induced brown adipogenic reprogramming of myoblasts. (D and E) qPCR analysis of general adipogenic genes (D) and brown adipogenic genes (E) in reprogrammed cells induced by Bex treatment, RXRα/β/γ overexpression of each subtype, or combined treatment (n = 3). (F) UCP1 immunostaining in reprogrammed brown adipogenic cells induced by Bex treatment and RXR overexpression. (G) Ucp1 expression in reprogrammed cells from primary myoblasts induced by Bex treatment, RXRα/β/γ subtype overexpression, or combined treatment (n = 3). Data were normalized to Ctrl and represent mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < (H) RXRα, RXRβ, and RXRγ mRNA levels in WATs, BAT, and skeletal muscle (SKM). mRNA was purified in WATs, BAT, and SKM from mice (n = 5). Values are normalized to the RXRα level in epiWAT and expressed as mean ± SEM. ∗p < 0.05, ∗∗∗p < Cell Reports  , DOI: ( /j.celrep ) Copyright © 2017 The Authors Terms and Conditions

5 Figure 3 Bex and RXR-Mediated Brown Adipogenic Reprogramming Is Only Partially Dependent on PRDM16 (A) Induction of Prdm16and Ucp1 expression by treatment with Bex and RXRα overexpression in myoblasts for the first several days. (B) Ucp1 and Prdm16 levels in Bex/RXR-treated C2C12 cells on day 2. Data were normalized to Ctrl and represent mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < (C) Knockdown of Prdm16 only partially reduced Bex/RXR-induced brown adipogenic reprogramming of myoblasts. Ucp1 and PPARγ were examined on day 6 (n = 3). (D) Bex and Prdm16 synergistically induced UCP1 expression in primary myoblasts. Data were normalized to Ctrl and represented mean ± SEM. ∗∗∗p < (n = 3). (E) Knockout of Prdm16 in C3H10T1/2 cells partially reduced the expression of Ucp1, Cox7a1, and Pparα and completely abolished Pgc1α expression but did not affect ap2 expression. Data were normalized to Ctrl and represent mean ± SEM (n = 3). Cell Reports  , DOI: ( /j.celrep ) Copyright © 2017 The Authors Terms and Conditions

6 Figure 4 Bex Initiates Browning Pathways at an Early Stage of Brown Adipogenic Reprogramming of Myoblasts (A) Scatterplot of differentially expressed genes in Ctrl and Bex-treated samples on day 2. Genes upregulated by Bex are marked in red, and those downregulated are shown in blue. (B) DAVID GO analysis of upregulated genes induced by Bex treatment. (C) Enriched molecular pathways of differentially expressed genes in GO-Elite. (D) Real-time PCR confirmed mRNA levels of some adipogenesis and browning genes induced by Bex in C2C12 cells on day 2. Data were normalized to Ctrl and represent mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < (n = 3). Cell Reports  , DOI: ( /j.celrep ) Copyright © 2017 The Authors Terms and Conditions

7 Figure 5 Bex Increases BAT Mass and Function In Vivo
(A) Bex treatment significantly reduced body weight gain in mice (n = 9). ∗p < 0.05; N.S., no significance. (B) Bex treatment increased oxygen consumption (VO2) in mice (n = 9). ∗p < 0.05. (C) Bex treatment increased heat release in mice (n = 9). ∗p < 0.05. (D) Bex treatment promoted cold tolerance (n = 9). ∗p < 0.05. (E) Bex increased the glucose tolerance test (GTT) and insulin tolerance test (ITT) response in mice (n = 9). ∗p < 0.05. (F) A positron emission tomography (PET) scan showed body composition in mice (n = 9). ∗p < 0.05. (G and H) Bex treatment increased BAT mass but decreased WAT mass in mice. Representative fat images (G) and quantification of different kinds of fat mass (H) (n = 9). ∗p < 0.05. (I) qPCR analysis of BAT genes in BAT, subWAT, epiWAT, and periWAT in control and Bex-treated mice (n = 9). Data were normalized to the vehicle group and represented mean ± SEM. ∗p < 0.05, ∗∗p < 0.01. (J) UCP1 immunostaining in BAT. Cell Reports  , DOI: ( /j.celrep ) Copyright © 2017 The Authors Terms and Conditions


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